自体骨膜包裹肌腱重建兔前交叉韧带对腱骨愈合影响的研究

发布时间：2018-03-25 21:00

  本文选题：前交叉韧带　切入点：自体肌腱　出处：《河北医科大学》2014年硕士论文

【摘要】：目的：观察自体骨膜包裹肌腱重建兔前交叉韧带（anterior cruciateligament，ACL）对腱骨愈合的影响，探讨自体骨膜促进腱骨愈合的能力与机制，为自体骨膜包裹肌腱在前交叉韧带损伤重建治疗领域的应用提供实验资料。 方法：选取96只3-4月龄新西兰大白兔，雌雄不限，体重2.5~3.5Kg，由河北医科大学实验动物中心提供。将实验用兔随机分为两组，每组48只，分别命名为A组、B组，，其中A组为实验组，B组为对照组。两组试验用兔饲养条件完全相同,术前麻醉用1%戊巴比妥钠溶液于耳缘静脉注射，剂量约为2ml/Kg，全麻后备皮消毒手术侧膝关节及远端部位，对两组新西兰大白兔分别进行直视下同侧自体跟腱移植重建前交叉韧带术,其中右膝关节做为手术侧。A组在进行自体跟腱移植重建前交叉韧带时取手术侧胫骨上端内侧骨膜，大小约1.5cm*0.5cm,用以包裹修整好的肌腱胫骨端，生发层朝向骨隧道，用可吸收缝线缝合固定，将骨膜包裹的肌腱植入胫骨隧道内，生发层与骨隧道紧密接触；B组肌腱无骨膜包裹，只进行自体跟腱移植前交叉韧带重建。术中给予庆大霉素4万单位预防感染,术后继续抗炎治疗3日,支具固定手术侧膝关节，常规换药,14天拆除缝线,分别于术后第2周、第4周、第6周、第8周麻醉后处死实验用兔,解剖手术侧膝关节,将实验用兔手术侧膝关节包含重建韧带隧道部分完整解剖并离断，剔除周围软组织，将胫骨隧道部位标本分为两段，其中一段置于4%甲醛溶液中固定,另一段置于2%戊二醛溶液中固定。将所需标本置于含30%甲酸溶液及10%甲醛溶液配置的脱钙液中进行脱钙48h后冲水48h。将4%甲醛溶液中固定的标本分别进行HE染色及甲苯胺蓝（toluidineblue,TB）染色后进行光镜制片，将2%戊二醛溶液中固定的标本进行电镜(transmission electron microscope,TEM)制片，制片完成后分别用光镜和电镜观察两组重建前交叉韧带腱骨愈合的情况。 结果： 1实验动物术后均未出现感染、伤口不愈合等并发症。 2新西兰白兔自体跟腱移植重建前交叉韧带后lachman试验阴性，侧方应力试验阴性，符合交叉韧带重建后的临床要求。 3大体观察：实验组自体跟腱移植重建前交叉韧带术后2周时隧道腱骨结不紧密，腱骨之间有疏松的结缔组织存在，外力作用下可导致腱骨分离；术后4周时隧道腱骨紧密程度好于2周时，腱骨之间有稍致密的结缔组织存在，外力作用下导致腱骨分离难于2周时；术后6周时隧道腱骨紧密程度好于4周时，腱骨之间有致密的结缔组织存在，外力作用下导致腱骨分离难于4周时；术后8周时隧道腱骨紧密程度好于6周时，腱骨之间有较致密的结缔组织存在，外力作用下不易导致腱骨分离。对照组自体跟腱移植重建前交叉韧带术后第2周、第4周、第6周、第8周时隧道腱骨结合程度均弱于同时期的实验组，腱骨之间的结缔组织均少于同期实验组，外力作用下较实验组更容易导致腱骨分离。 4显微镜下观察：实验组第2周时腱骨结合部有大量细胞浸润，主要为炎性细胞，其次为成纤维细胞，并出现少量纤维软骨细胞，HE染色未见明显着色；术后第4周、第6周及第8周可见损伤裂口处新生组织填充逐渐增多，并与周围肌腱及骨隧道组织形成紧密连接，新生组织中纤维软骨细胞逐渐增多，细胞外基质成分逐渐增多并出现纤维成分及新生毛细血管形成；甲苯胺蓝染色范围逐渐扩大，表明新生组织中软骨成分逐渐增多。对照组腱骨结合部2周，少量细胞浸润，肌腱与骨隧道未紧密结合，亦无纤维软骨细胞出现；术后4周、6周及8周可见腱骨结合部出现少量新生组织，但与周围组织连接不紧密，其中的纤维软骨细胞数量明显少于实验组，仅有少量新生毛细血管形成，细胞基质成分较少，甲苯胺蓝染色见着色不明显(FIG1.1-4.8)。统计学分析结果显HE染色及甲苯胺蓝（TB）染色术后第2周、第4周、第6周同时期实验组和对照组组间比较无明显差异，术后第8周实验组和对照组组间比较有统计学意义（P＜0.05）。术后第2周至第8周实验组组内比较有统计学意义，术后第2周至第8周对照组组内比较有统计学意义（P＜0.05）。电镜观察组术后第6周及第8周组间比较有统计学意义（P＜0.05）。(Table-7) 结论： 骨膜包裹自体跟腱移植重建兔前交叉韧能够促进腱骨结合部炎性反应发生和细胞聚集，促进炎性细胞，成纤维细胞、骨细胞及软骨细胞的再生；促进新生毛细血管的形成及软骨分化；对腱骨结合部愈合修复具有积极的促进作用。
[Abstract]:Objective: To observe the effect of autologous periosteum wrapped tendon reconstruction of anterior cruciate ligament in rabbits (anterior cruciateligament, ACL) on tendon bone healing, to explore the ability and mechanism of autologous periosteum promote tendon bone healing and provide experimental data for the application of autologous periosteum wrapped tendon reconstruction for the treatment in the field of anterior cruciate ligament injury.
Methods: a total of 96 3-4 month old New Zealand white rabbits, male or female, weight 2.5~3.5Kg, provided by the experimental animal center of Hebei Medical University. The experimental rabbits were randomly divided into two groups, 48 rats in each group, which were named as A group, B group, A group for the experimental group, group B as control group. Two groups test with rabbit under the same feeding conditions, anesthesia by ear vein injection of 1% pentobarbital sodium before operation, the dose is about 2ml/Kg, the skin is disinfected after general anesthesia in surgical knee joint and distal part of two groups of New Zealand white rabbits were reconstructed at ipsilateral autologous tendon anterior cruciate ligament, the right knee to do for the surgical side group.A cruciate ligament reconstruction in self tendon before surgery from the proximal tibia medial periosteum, the size of about 1.5cm*0.5cm, used to wrap the trimmed tibial tendon, the germinal layer toward the bone tunnel, with absorbable suture. That will be the periosteum wrapped tendon implanted into the tibial tunnel within the germinal layer in close contact with the bone tunnel; B group without periosteum wrapped tendon, only self tendon anterior cruciate ligament reconstruction. Given gentamycin 40 thousand units to prevent infection during operation, postoperative anti-inflammatory treatment for 3 days, with a fixed surgical knee joint, conventional 14 days of dressing, suture, respectively for second weeks, after fourth weeks, sixth weeks, eighth weeks after anesthesia were experimental rabbit knee joint surgery, anatomy, the experimental rabbit knee joint ligament reconstruction surgery includes complete transection and anatomic section of the tunnel, excluding the surrounding soft tissue. The tibial tunnel position divided into two sections, one fixed segment in the 4% Formaldehyde Solution, another fixed in 2% glutaraldehyde solution. The decalcified specimens were placed in liquid containing 30% formic acid and 10% Formaldehyde Solution in the 48h configuration after decalcification water 48h. 4% formaldehyde solution Liquid fixed specimens were stained with HE and toluidine blue respectively (toluidineblue, TB) after staining by light microscope method, were fixed in 2% glutaraldehyde solution for the electron microscope (transmission electron microscope, TEM) production, production is completed respectively by light microscope and electron microscopy observation cross tendon bone healing of two groups before reconstruction.
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